Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2015 |
| Outros Autores: | , , , , |
| Tipo de documento: | Artigo |
| Idioma: | por |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/23244 |
Resumo: | Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Recife, PE, Brasil |
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Mariz, F. C.Coimbra, E. C.Jesus, A. L. S.Nascimento, L. M.Torres, F. A. G.Freitas, A. C.2017-11-16T18:12:49Z2017-11-16T18:12:49Z2015MARIZ, F. C. et al. Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter. BioMed Research International, v. 2015, p. 594120, 2015.2314-6141https://www.arca.fiocruz.br/handle/icict/2324410.1155/2015/594120porBiotecnologiaProteínas do CapsídeoProteínas Oncogênicas ViraisNeoplasias do Colo do ÚteroVacinas de Partículas Semelhantes a VírusBiotechnologyCapsid ProteinsFemaleGene Expression Regulation, ViralHuman papillomavirus 16HumansOncogene Proteins, ViralPichiaPromoter Regions, GeneticUterine Cervical NeoplasmsVaccines, Virus-Like ParticleBiotecnologiaProteínas do CapsídeoimunologiaProteínas Oncogênicas ViraisimunologiaNeoplasias do Colo do Úteroprevenção & controleVacinas de Partículas Semelhantes a VírusFemininoRegulação Viral da Expressão GênicaPapillomavirus Humano 16patogenicidadeHumanosquímicaPichiaRegiões Promotoras GenéticasvirologiaDevelopment of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoterinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Aggeu Magalhães. Recife, PE, BrasilThe human papillomavirus (HPV) L1 major capsid protein, which forms the basis of the currently available vaccines against cervical cancer, self-assembles into virus-like particles (VLPs) when expressed heterologously. We report the development of a biotechnology platform for HPV16 L1 protein expression based on the constitutive PGK1 promoter (PPGK1) from the methylotrophic yeast Pichia pastoris. The L1 gene was cloned under regulation of PPGK1 into pPGKΔ3 expression vector to achieve intracellular expression. In parallel, secretion of the L1 protein was obtained through the use of an alternative vector called pPGKΔ3α, in which a codon optimized α-factor signal sequence was inserted. We devised a work-flow based on the detection of the L1 protein by dot blot, colony blot, and western blot to classify the positive clones. Finally, intracellular HPV VLPs assembly was demonstrated for the first time in yeast cells. This study opens up perspectives for the establishment of an innovative platform for the production of HPV VLPs or other viral antigens for vaccination purposes, based on constitutive expression in P. pastoris.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/23244/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINAL26090426 2015 mar-dev.oa.pdf26090426 2015 mar-dev.oa.pdfapplication/pdf8169612https://www.arca.fiocruz.br/bitstream/icict/23244/2/26090426%202015%20mar-dev.oa.pdf061d9682d07ebcf75d4e5174d193f4fdMD52TEXT26090426 2015 mar-dev.oa.pdf.txt26090426 2015 mar-dev.oa.pdf.txtExtracted texttext/plain54776https://www.arca.fiocruz.br/bitstream/icict/23244/3/26090426%202015%20mar-dev.oa.pdf.txt80f6420117d67db7578a61ebccf0cab1MD53icict/232442018-08-15 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| dc.title.pt_BR.fl_str_mv |
Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter |
| title |
Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter |
| spellingShingle |
Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter Mariz, F. C. Biotecnologia Proteínas do Capsídeo Proteínas Oncogênicas Virais Neoplasias do Colo do Útero Vacinas de Partículas Semelhantes a Vírus Biotechnology Capsid Proteins Female Gene Expression Regulation, Viral Human papillomavirus 16 Humans Oncogene Proteins, Viral Pichia Promoter Regions, Genetic Uterine Cervical Neoplasms Vaccines, Virus-Like Particle Biotecnologia Proteínas do Capsídeo imunologia Proteínas Oncogênicas Virais imunologia Neoplasias do Colo do Útero prevenção & controle Vacinas de Partículas Semelhantes a Vírus Feminino Regulação Viral da Expressão Gênica Papillomavirus Humano 16 patogenicidade Humanos química Pichia Regiões Promotoras Genéticas virologia |
| title_short |
Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter |
| title_full |
Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter |
| title_fullStr |
Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter |
| title_full_unstemmed |
Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter |
| title_sort |
Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter |
| author |
Mariz, F. C. |
| author_facet |
Mariz, F. C. Coimbra, E. C. Jesus, A. L. S. Nascimento, L. M. Torres, F. A. G. Freitas, A. C. |
| author_role |
author |
| author2 |
Coimbra, E. C. Jesus, A. L. S. Nascimento, L. M. Torres, F. A. G. Freitas, A. C. |
| author2_role |
author author author author author |
| dc.contributor.author.fl_str_mv |
Mariz, F. C. Coimbra, E. C. Jesus, A. L. S. Nascimento, L. M. Torres, F. A. G. Freitas, A. C. |
| dc.subject.other.pt_BR.fl_str_mv |
Biotecnologia Proteínas do Capsídeo Proteínas Oncogênicas Virais Neoplasias do Colo do Útero Vacinas de Partículas Semelhantes a Vírus |
| topic |
Biotecnologia Proteínas do Capsídeo Proteínas Oncogênicas Virais Neoplasias do Colo do Útero Vacinas de Partículas Semelhantes a Vírus Biotechnology Capsid Proteins Female Gene Expression Regulation, Viral Human papillomavirus 16 Humans Oncogene Proteins, Viral Pichia Promoter Regions, Genetic Uterine Cervical Neoplasms Vaccines, Virus-Like Particle Biotecnologia Proteínas do Capsídeo imunologia Proteínas Oncogênicas Virais imunologia Neoplasias do Colo do Útero prevenção & controle Vacinas de Partículas Semelhantes a Vírus Feminino Regulação Viral da Expressão Gênica Papillomavirus Humano 16 patogenicidade Humanos química Pichia Regiões Promotoras Genéticas virologia |
| dc.subject.en.pt_BR.fl_str_mv |
Biotechnology Capsid Proteins Female Gene Expression Regulation, Viral Human papillomavirus 16 Humans Oncogene Proteins, Viral Pichia Promoter Regions, Genetic Uterine Cervical Neoplasms Vaccines, Virus-Like Particle |
| dc.subject.decs.pt_BR.fl_str_mv |
Biotecnologia Proteínas do Capsídeo imunologia Proteínas Oncogênicas Virais imunologia Neoplasias do Colo do Útero prevenção & controle Vacinas de Partículas Semelhantes a Vírus Feminino Regulação Viral da Expressão Gênica Papillomavirus Humano 16 patogenicidade Humanos química Pichia Regiões Promotoras Genéticas virologia |
| description |
Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Recife, PE, Brasil |
| publishDate |
2015 |
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2015 |
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2017-11-16T18:12:49Z |
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2017-11-16T18:12:49Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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MARIZ, F. C. et al. Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter. BioMed Research International, v. 2015, p. 594120, 2015. |
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https://www.arca.fiocruz.br/handle/icict/23244 |
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2314-6141 |
| dc.identifier.doi.none.fl_str_mv |
10.1155/2015/594120 |
| identifier_str_mv |
MARIZ, F. C. et al. Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter. BioMed Research International, v. 2015, p. 594120, 2015. 2314-6141 10.1155/2015/594120 |
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